LOCUS       sequence-415894-        8929 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of rat alpha2delta-1 calcium channel auxillary subunit 
            with a single HA tag inserted after aa549 and with an R421A mutation
            that disrupts Gabapentin binding.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8929)
  AUTHORS   Cassidy JS, Ferron L, Kadurin I, Pratt WS, Dolphin AC
  TITLE     Functional exofacially tagged N-type calcium channels elucidate the 
            interaction with auxiliary alpha2delta-1 subunits.
  JOURNAL   Proc Natl Acad Sci U S A. 2014 Jun 17;111(24):8979-84. doi: 
            10.1073/pnas.1403731111. Epub 2014 Jun 2.
  PUBMED    24889613
REFERENCE   2  (bases 1 to 8929)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1073/pnas.1403731111"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2014-06-17- 17"; volume: "111"; issue: "24"; pages: "8979-84"
COMMENT     Sequence Label: cacna2d1 R241A HA pMT2
FEATURES             Location/Qualifiers
     source          1..8929
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             90..653
                     /codon_start=1
                     /gene="Mus musculus Dhfr"
                     /product="mouse dihydrofolate reductase"
                     /label=DHFR
                     /translation="MVRPLNCIVAVSQNMGIGKNGDLPWPPLRNEFKYFQRMTTTSSVE
                     GKQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRELKEPPRGAHFLAKSLDDALRLIEQP
                     ELASKVDMVWIVGGSSVYQEAMNQPGHLRLFVTRIMQEFESDTFFPEIDLGKYKLLPEY
                     PGVLSEVQEEKGIKYKFEVYEKKD"
     polyA_signal    840..974
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(877..896)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     931..950
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1647..1664)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1818..2406)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1898..1917)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2577..3368,3369..3437))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3200..3219
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3438..3542)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3610..3628
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3666..3688)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3788..3807
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4001..4023
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4015..4032
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4016..4032
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(4042..4371)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4056..4191
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4356..4376
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     intron          4840..5069
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     5052..5071
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             6826..6852
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
ORIGIN
        1 gggggggaca gctcagggct gcgatttcgc gccaaacttg acggcaatcc tagcgtgaag
       61 gctggtagga ttttatcccc gctgccatca tggttcgacc attgaactgc atcgtcgccg
      121 tgtcccaaaa tatggggatt ggcaagaacg gagacctacc ctggcctccg ctcaggaacg
      181 agttcaagta cttccaaaga atgaccacaa cctcttcagt ggaaggtaaa cagaatctgg
      241 tgattatggg taggaaaacc tggttctcca ttcctgagaa gaatcgacct ttaaaggaca
      301 gaattaatat agttctcagt agagaactca aagaaccacc acgaggagct cattttcttg
      361 ccaaaagttt ggatgatgcc ttaagactta ttgaacaacc ggaattggca agtaaagtag
      421 acatggtttg gatagtcgga ggcagttctg tttaccagga agccatgaat caaccaggcc
      481 acctcagact ctttgtgaca aggatcatgc aggaatttga aagtgacacg tttttcccag
      541 aaattgattt ggggaaatat aaacttctcc cagaataccc aggcgtcctc tctgaggtcc
      601 aggaggaaaa aggcatcaag tataagtttg aagtctacga gaagaaagac taacaggaag
      661 atgctttcaa gttctctgct cccctcctaa agctatgcat ttttataaga ccatgggact
      721 tttgctggct ttagatcata atcagccata ccacatttgt agaggtttta cttgctttaa
      781 aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta
      841 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
      901 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
      961 atcatgtctg gatccccggc caacggtctg gtgacccggc tgcgagagct cggtgtacct
     1021 gagacgcgag taagcccttg agtcaaagac gtagtcgttg caagtccgca ccaggtactg
     1081 atatcccacc aaaaagtgcg gcggcggctg gcggtagagg ggccagcgta gggtggccgg
     1141 ggctccgggg gcgaggtctt ccaacataag gcgatgatat ccgtagatgt acctggacat
     1201 ccaggtgatg ccggcggcgg tggtggaggc gcgcggaaag tcgcggacgc ggttccagat
     1261 gttgcgcagc ggcaaaaagt gctccatggt cgggacgctc tggccggtga ggcgtgcgca
     1321 gtcgttgacg ctctagaccg tgcaaaagga gagcctgtaa gcgggcactc ttccgtggtc
     1381 tggtggataa attcgcaagg gtatcatggc ggacgaccgg ggttcgaacc ccggatccgg
     1441 ccgtccgccg tgatccatgc ggttaccgcc cgcgtgtcga acccaggtgt gcgacgtcag
     1501 acaacggggg agcgctcctt ttggcttcct tccaggcgcg gcggctgctg cgctagcttt
     1561 tttggcgagc tcgaattaat tctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     1621 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     1681 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     1741 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     1801 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     1861 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     1921 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     1981 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     2041 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     2101 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     2161 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     2221 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     2281 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     2341 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     2401 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     2461 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     2521 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     2581 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     2641 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     2701 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     2761 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     2821 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     2881 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     2941 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     3001 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     3061 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     3121 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     3181 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     3241 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     3301 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     3361 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     3421 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     3481 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     3541 cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta
     3601 acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt
     3661 gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc
     3721 gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt
     3781 aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg
     3841 cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag gctgcgcaac
     3901 tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga
     3961 tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa
     4021 acgacggcca gtgccaagct ttttgcaaaa gcctaggcct ccaaaaaagc ctcctcacta
     4081 cttctggaat agctcagagg ccgaggcggc ctcggcctct gcataaataa aaaaaattag
     4141 tcagccatgg ggcggagaat gggcggaact gggcggagtt aggggcggga tgggcggagt
     4201 taggggcggg actatggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc
     4261 tggggagcct ggggactttc cacacctggt tgctgactaa ttgagatgca tgctttgcat
     4321 acttctgcct gctggggagc ctggggactt tccacaccct aactgacaca cattccacag
     4381 gatccggtcg cgcgaatttc gagcggtgtt ccgcggtcct cctcgtatag aaactcggac
     4441 cactctgaga cgaaggctcg cgtccaggcc agcacgaagg aggctaagtg ggaggggtag
     4501 cggtcgttgt ccactagggg gtccactcgc tccagggtgt gaagacacat gtcgccctct
     4561 tcggcatcaa ggaaggtgat tggtttatag gtgtaggcca cgtgaccggg tgttcctgaa
     4621 ggggggctat aaaagggggt gggggcgcgt tcgtcctcac tctcttccgc atcgctgtct
     4681 gcgagggcca gctgttgggc tcgcggttga ggacaaactc ttcgcggtct ttccagtact
     4741 cttggatcgg aaacccgtcg gcctccgaac ggtactccgc caccgaggga cctgagcgag
     4801 tccgcatcga ccggatcgga aaacctctcg actgttgggg tgagtactcc ctctcaaaag
     4861 cgggcatgac ttctgcgcta agattgtcag tttccaaaaa cgaggaggat ttgatattca
     4921 cctggcccgc ggtgatgcct ttgagggtgg ccgcgtccat ctggtcagaa aagacaatct
     4981 ttttgttgtc aagcttgagg tgtggcaggc ttgagatctg gccatacact tgagtgacaa
     5041 tgacatccac tttgcctttc tctccacagg tgtccactcc caggtccaac tgcaggcgag
     5101 cctgaattcc tcgagggtac cgagctcgga tccactagta acggccgcca gtgtgctgga
     5161 attctgcaga tcgcgaagat ggctgctggc tgcctgctgg ccttgactct gacacttttc
     5221 caatcttggc tgatcggccc ctcgagcgag gagcccttcc cttcgcccgt cactatcaag
     5281 tcatgggtgg ataaaatgca ggaagacctt gtcacactgg caaagacagc aagtggagtg
     5341 actcagcttg ctgacattta tgaaaaatac caagatttgt atactgtgga gccaaataat
     5401 gcacgccaac tggttgaaat tgcagctaga gacattgaga aacttctgag caacagatct
     5461 aaagccctgg tgcgcctggc tatggaagca gagaaagtcc aagccgccca ccaatggagg
     5521 gaagattttg caagcaacga agttgtctac tacaacgcta aagacgatct tgatcctgaa
     5581 agaaatgaga gtgagtcagg cagccaacgg attaaacctg ttttcatcga agacgctaat
     5641 tttggacgcc agatatccta tcaacatgca gcggtccata ttcccacgga catctatgag
     5701 ggctcaacca tagtgttaaa cgaactcaac tggacaagtg ccttagatga agttttcaaa
     5761 agaaatcgag atgaagaccc tacattgctg tggcaagtgt ttggcagcgc caccggcctg
     5821 gcccgatatt atccagcttc tccgtgggtg gataacagca gaactccaaa caagatcgat
     5881 ctctacgatg tacgcagagc accatggtac atccagggag ctgcatcccc caaagacatg
     5941 ctcattctgg tggacgtgag tggaagtgtg agcggattga ctctgaaact catccggaca
     6001 tctgtctctg agatgttaga aaccctttcc gatgatgatt tcgtgaatgt agcttcattt
     6061 aacagcaatg ctcaggatgt gagttgtttc cagcacctgg ttcaagcgaa tgtaaggaac
     6121 aagaaggtgc tgaaggatgc ggtgaataac atcacagcaa aggggataac agattacaag
     6181 aaaggcttta gcttcgcgtt tgaacagcta cttaattata atgtttccag agccaattgt
     6241 aataagatca tcatgttatt cacggatgga ggagaagaga gagcccagga gatatttgcc
     6301 aagtacaata aagacaaaaa agtccgcgtg tttacatttt ctgttggtca acataattat
     6361 gacagaggac ctattcagtg gatggcttgt gaaaacaaag gttattatta tgagattccc
     6421 tctattggtg caataagaat caatactcag gaatatctag atgttctggg aagaccaatg
     6481 gttttagccg gggacaaagc aaagcaagtc cagtggacaa atgtgtattt ggatgctctg
     6541 gagctgggac ttgtcattac tggaactcta ccggtcttca acgtcactgg ccagtctgaa
     6601 aataagacta acttgaagaa ccagttgatt cttggtgtga tgggcgttga tgtgtctctg
     6661 gaagatatta agagattgac accacgtttt acactgtgcc ccaacggtta ctattttgca
     6721 atcgatccta atggttatgt cttactgcat ccaaatcttc agccaaagaa ccccaaatct
     6781 caggagccgg tcacactgga ttttctcgat gctgagttag agaattaccc ttacgacgtc
     6841 cctgactacg ccgatattaa agtggagatt cgaaataaaa tgatagacgg agaaagtgga
     6901 gaaaaaacat tcagaactct ggtcaaatct caagatgaga gatatattga caaaggaaat
     6961 cgaacataca catggacacc tgtcaatggc acagattaca gtttggcctt ggtattgcca
     7021 acctacagtt tttactatat aaaagccaaa atagaagaga caataactca ggccagatca
     7081 aaaaagggga aaatgaagga ttcagaaacc ctgaagccag acaattttga agaatctggc
     7141 tacactttca tagcaccaag ggaatactgc aatgacctga aaccttcaga taataacact
     7201 gaatttcttt taaatttcaa tgaatttatt gataggaaaa ctccaaacaa cccttcctgt
     7261 aatacagatt tgattaatag aatcttgctg gatgcgggtt ttacaaatga acttgtccaa
     7321 aattattgga gtaaacagaa aaatatcaaa ggagtgaagg cacgctttgt ggtgactgat
     7381 ggcggaatta caagagttta ccccaaagag gctggagaaa actggcaaga gaacccagag
     7441 acgtatgagg acagcttcta caaacggagc ctagataacg ataactacgt tttcactgcg
     7501 ccctacttca acaaaagtgg acctggtgcc tatgaatctg gaattatggt aagcaaagct
     7561 gtagaactgt acatccaagg aaaacttctt aagcctgcag ttgtgggaat taaaattgac
     7621 gtaaactcct ggatagaaaa ttttaccaaa acttcaatca gggatccgtg tgctggtcca
     7681 gtttgtgact gcaaaagaaa cagtgatgta atggactgtg tcattctaga tgatggtgga
     7741 tttcttctga tggccaatca tgatgattac actaatcaga ttggacgttt ttttggagag
     7801 attgacccga gcatgatgag acacctggtt aatatatcac tttatgcatt caataaatca
     7861 tatgactatc agtctgtgtg tgatccaggg gcagcaccaa agcaaggagc aggacatcgc
     7921 tcagcctatg tgccatcaat tacagacata ctccagattg gctggtgggc cactgctgcc
     7981 gcctggtcta ttctccagca gctactcttg agtttgacat ttccacggct ccttgaggca
     8041 gttgaaatgg aagaggatga cttcacagct tccctgtcta agcagagctg catcacagaa
     8101 caaactcagt acttcttcaa gaacgatact aaatcattca gtggtttact ggactgtgga
     8161 aactgttcca ggatctttca tgttgagaag cttatgaaca ccaacctagt gttcataatg
     8221 gtggagagca agggaacatg tccgtgtgac acgcggctgc tcatgcaagc agaacagact
     8281 tctgatggtc cagatccttg cgacatggtt aagcagccca gataccgaaa aggacctgat
     8341 gtctgctttg ataacaatgt gctggaggat tataccgact gtggtggtgt ttctggatta
     8401 aacccttcct tatggtctat ctttggactc caatttatac tcctttggct ggtatctggc
     8461 agcagacact atctatggtg accctctaaa cccatgatct gcaacttaat tccatgccct
     8521 gccaaaacat gagccctcag ttacattaaa atagagcccg ttgaagaagc tccacagaac
     8581 cttagctggg cctctaccct gatgtaactg tgaggcttag attcctgacc atcgctggct
     8641 gcatgtcagg gtgttggaaa ctgaaactcg tgtgaacatg ctgcatcatc tgtgtccaac
     8701 atcaaagcaa agctctactc attctctgtg ggaggtttgg gagttcgctg ttgcactgtt
     8761 ggagattacg tggaaaaggc tcccccacgc aggccttatg aaaagaagca gtcttgactt
     8821 tgacctagca tttcaacctg ctgcagtttt accaagccga attcctgcag cccgggggat
     8881 ccactagatc catcacactg gcggccgcgt cgacgaattc cgggggggg
//
